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Efficient Methods to Generate Reproducible Mass Spectra in Matrix-Assisted Laser Desorption Ionization of Peptides

  • Research Article
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Journal of The American Society for Mass Spectrometry

Abstract

In our previous matrix-assisted laser desorption ionization (MALDI) studies of peptides, we found that their mass spectra were virtually determined by the effective temperature in the early matrix plume, Tearly, when samples were rather homogeneous. This empirical rule allowed acquisition of quantitatively reproducible spectra. A difficulty in utilizing this rule was the complicated spectral treatment needed to get Tearly. In this work, we found another empirical rule that the total number of particles hitting the detector, or TIC, was a good measure of the spectral temperature and, hence, selection of spectra with the same TIC resulted in reproducible spectra. We also succeeded in obtaining reproducible spectra throughout a measurement by controlling TIC near a preset value through feedback adjustment of laser pulse energy. Both TIC selection and TIC control substantially reduced the shot-to-shot spectral variation in a spot, spot-to-spot variation in a sample, and even sample-to-sample variation in MALDI using α-cyano-4-hydroxycinnamic acid or 2,5-dihydroxybenzoic acid as matrix. Based on the utilization of acquired data, TIC control was more efficient than TIC selection by an order of magnitude. Both techniques produced calibration curves with excellent linearity, suggesting their utility in quantification of peptides.

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Acknowledgments

The authors acknowledge support for this work by the National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST) (2012054350). S.H.A. and K.M.P. thank the Ministry of Education, Science, and Technology, Republic of Korea, for Brain Korea 21 Fellowship.

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Authors and Affiliations

  1. Department of Chemistry, Seoul National University, Seoul, 151-742, Korea

    Sung Hee Ahn, Kyung Man Park, Yong Jin Bae & Myung Soo Kim

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  1. Sung Hee Ahn
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  2. Kyung Man Park
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  3. Yong Jin Bae
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  4. Myung Soo Kim
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Correspondence to Myung Soo Kim.

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Ahn, S.H., Park, K.M., Bae, Y.J. et al. Efficient Methods to Generate Reproducible Mass Spectra in Matrix-Assisted Laser Desorption Ionization of Peptides. J. Am. Soc. Mass Spectrom. 24, 868–876 (2013). https://doi.org/10.1007/s13361-013-0612-2

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  • DOI: https://doi.org/10.1007/s13361-013-0612-2

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